Anti-convulsant

ABSTRACT

New amino-benzamides and their use for the treatment of epilepsy.

BACKGROUND OF THE INVENTION

1. Field of the Invention

This invention relates to a new type of anti-convulsant drug. Specifically, it deals with new benzamide compounds and their use for the treatment of epilepsy.

2. Description of the Prior Art

In the treatment of epilepsy or other convulsing causing diseases, various drugs, such as Phenobarbital, Phenytoin, Mephenytoin, and Carbamazapine have been used to prevent their attacks. These drugs, however, have undesirable side-effects in most cases.

There is, therefore, a need for a new type of anti-convulsant drug which will have fewer side-effects than those presently used.

BRIEF SUMMARY OF THE INVENTION

It is, accordingly, one object of the present invention to set forth a new anti-convulsant drug.

Another object of the present invention is to set forth new drugs of the amino-benzamide type for the treatment of epilepsy which have fewer side-effects.

A still further object of the present invention is to set forth new amino-benzamide anti-convulsant drugs and method for using the same for the treatment of epilepsy.

These and other advantages of the present invention will be apparent from the following detailed description and test data.

In accordance with the above objects, it has been found that amino-benzamide compounds are capable of preventing convulsing attacks caused by epilepsy. These compounds are various ortho, meta, and para amino-benzamides. These compounds are:

1. 4-amino-N-phenyl-benzamide; [4-aminobenzanilide]

2. 4-amino-N-cyclohexyl-benzamide

3. 4-amino-N-amyl-benzamide; [4-amino-N-(n-pentyl)-benzamide]

4. 4-amino-N-benzyl-benzamide

5. 4-amino-N,N-(di-n-propyl)-benzamide

6. 4-amino-N-(n-hexyl)-benzamide

7. 4-amino-N-(n-butyl)-benzamide

8. 4-amino-N-(N-methylbenzyl)-benzamide ##STR1## 11. 4-amino-N-(alpha-methylbenzyl)-benzamide

With the drugs available today, complete control of epilepsy can be achieved in about 60% of epilepsy cases. These amino-benzamides represent a new class of compounds for the treatment of epilepsy. The highly potent anti-convulsant activity of 4-amino-N-(alpha-methylbenzyl)-benzamide offers some unique advantages over currently available drugs. In animal studies, this amino-benzamide is effective against a wide range of convulsion types. This activity range is broader than many of the currently available drugs. Additionally, the amide offers a wide margin of safety between the dose required to produce anti-convulsant effects and the dose which produces any central nervous system side-effects. These facts all indicate that the amino-benzamides have the potential to improve the current drug therapy for epilepsy.

DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

The following equations illustrate the formation of the benzamide compounds of the present invention and their use using 4-amino-N-(alpha-methylbenzyl)-benzamide as an example: ##STR2##

The initial step (reaction A) involves the formation of the amide bond and the final reaction (B) is the conversion of the aromatic nitro-group to the desired amino-group.

Reaction A

A 1 L. 3-necked flask was equipped with a magnetic stir bar, reflux condensor with a drying tube, a pressure equalizing addition funnel, and a heating mantle. A 200-ml portion of 20% aqueous potassium carbonate was added to the flask followed by 10 g (0.083 mole) of d,l-alpha-methylbenzylamine (#2) in 200-ml of tetrahydrofuran. The resulting mixture was stirred and a solution of 23.3 g (0.125 mole) of 4-nitrobenzoylchloride (#1) in 100-ml of tetrahydrofuran was added dropwise from the addition funnel. Upon completion of the addition, the reaction mixture was heated to reflux for 12 hours then cooled and extracted 3 times with 100-ml portions of chloroform. The chloroform extracts were combined, dried over magnesium sulfate and evaporated to yield the desired compound, 4-nitro-N(alpha-methylbenzyl)-benzamide (#3). Recrystallization from benzene produced the purified compound.

Reaction B

A solution of 5 g (0.018 mole) of 4-nitro-N(alpha-methylbenzyl)-benzamide (#3) in 200-ml of absolute ethanol was added to a Paar hydrogenation bottle. To the solution was added a 0.4 g sample of 5% palladium on charcoal and the bottle placed on a Paar low-pressure hydrogenation apparatus. The mixture was shaken at room temperature in the presence of 45 p.s.i. of hydrogen for 2.5 hours. The catalyst was then removed by filtration through celite and the ethanol evaporated to yield the desired compound, 4-amino-N-(alpha-methylbenzyl)-benzamide (#4). Recrystallization from benzene produced the compound in pure form.

ADMINISTRATION

The anti-convulsant amide (4) was administered to mice and rats as a solution. The solution was prepared by dissolving the compound in 30% polyethylene glycol 400. The compound was injected either orally or intraperitoneally in a volume of 0.01 ml/g body weight in mice and 0.04 ml/10 g body weight in rats.

The amide is the only compound dissolved in the polyethylene glycol 400 solvent, thus the anti-convulsant activities observed do not depend on specific proportions of components. The use of the compound as a solution is only for ease of administration to animals and the activity observed is not based on the fact that the compound is in a solution. This is further pointed out by the fact that the solvent (30% polyethylene glycol 400) alone was used as a control in these studies (i.e., administration of the same volumes of solvent only produced no anti-convulsant effects).

ANTI-CONVULSANT ACTIVITY

The anti-convulsant activity of this compound (4-amino-N-(alpha-methylbenzyl)-benzamide) is based on results obtained from testing procedures in mice and rats. The anti-convulsant amide was compared with four prototype anti-epileptic agents (phenobarbital, phenytoin, mephenytoin and carbamazepine). The profile of anti-convulsant activity for each substance was established by five tests: one electrical and four chemical. The electrical test employed was the maximal electroshock seizure (MES) pattern test. The four chemical tests included the subcutaneous Metrazol Seizure Threshold Test (sc Met), subcutaneous Bicuculline Seizure Threshold Test (sc Bic), subcutaneous Picrotoxin Seizure Threshold Test (sc Pic), and the subcutaneous Strychnine Seizure Pattern Test (sc Strych). These tests are conducted by administering the anti-convulsant compound or drug to the animals then giving the convulsive amount of electrical energy or chemical convulsant and measuring the ability of the test compound to prevent the production of convulsions.

To determine anti-convulsant potency and toxicity, groups of at least eight mice were tested with various doses of the drug until at least four points were established between the limits of 100% protection or toxicity and 0% protection or toxicity. The dose of drug required to produce the desired endpoint in 50% of animals (ED50), 95% confidence interval, slope of the regression line, and the standard error of the slope were then calculated.

The activity data comparing compound 4 with the four prototype anti-convulsant drugs are given in the following Tables. These data were obtained in testing conducted by the Anticonvulsant Drug Development Program, Epilepsy Branch, Neurological Disorders Program, National Institute of Neurological and Communicative Disorders and Stroke. The tests used are described in J. F. Reinhard and J. F. Reinhard, Jr., "Experimental Evaluation of Anticonvulsants," in Anticonvulsants, J. A. Vida, Ed., Academic Press, New York, N.Y., 1977.

In order to better understand the terminology in the tables, the following definitions of terms therein are set forth.

Description of Data Terms

(a) MES=Maximal electroshock seizure test. This test measures the ability of the test drug to abolish the hind-limb tonic-extensor component of maximal seizures induced by 50 mA of current delivered to the animals for 0.2 seconds.

(b) scMet=Subcutaneous metrazol seizure threshold test. The test measures the ability of the test drug to afford complete protection against a subcutaneously administered convulsant dose of metrazol. Metrazol is a very potent CNS stimulant which in sufficient doses produces convulsions.

(c) scBic=Subcutaneous bicuculline seizure threshold test. Explanation same as in (b).

(d) scPic=Subcutaneous picrotoxin seizure threshold test. Explanation same as in (b)

(e) ScStrych=Subcutaneous strychnine seizure pattern test. The test measures the ability of the test drug to abolish all toxic components of seizures induced by the subcutaneous injection of a convulsant dose of strychnine.

(f) ED50=Effective dose in 50% of the subjects (median effective dose), the dose of the drug in mg/kg of animal weight required to produce the pharmacological effect in 50% of the animals in a group.

(g) TD50=Toxic dose in 50% of the subjects, dose required to produce toxic side-effects in 50% of the animals tested. The test for toxic effects is referred to as the "Rotorod Toxicity Test." The test shows the loss of an animal's ability to maintain its balance on a rotating rod. Untreated or normal animals can easily maintain their balance in this test.

(h) HD50=Hypnotic dose in 50% of the subjects tested. Hypnosis is defined as loss of righting reflex--when placed on its back, the animal does not have the ability to get to its feet.

(i) LD50=Lethal dose in 50% of the subjects, the dose required to kill 50% of the animals.

(j) PI=Protective index, defined as the ratio of TD50 to ED50 (TD50/ED50). The PI is a measure of the safety of the drug. A high PI value indicates a greater margin of safety between the effective dose for the pharmacological effect and the toxic dose producing side-effects. A compound with a high PI would be a safer drug to use.

(k) 95% Confidence Interval--This is a statistical term which says that if we repeated a given test a number of times, 95% of the time the result would lie within the limits defined as the 95% confidence interval. Example: From Table 1 the ED50 for 4-amino-N-(alpha-methylbenzyl)-benzamide in the MES test is given as 18.02 mg/kg. If we were to repeat this test 100 times, the ED50 which we would determine would fall between 13.41-21.43 mg/kg 95% of the time. The 95% confidence interval then represents a range of doses.

(l) Slope of regression line--This represents the rate of change in response (effect) for a change in dose. The higher the numerical value of the slope, the greater the rate of change in response.

                                      TABLE 1.                                     __________________________________________________________________________     Profile of Anti-convulsant Activity of                                         Intraperitoneally Administered 4-amino-N--                                     (alpha-methylbenzyl)-benzamide and Some Proto-                                 type Anti-epileptic Drugs in Mice                                                     Time                                                                           of                                                                      ADD No.                                                                               Test                                                                              Rotorod ED50 (mg/kg) and PI                                          Substance                                                                             (hrs)                                                                             TD50 (mg/kg)                                                                           MES       sc Met                                             __________________________________________________________________________     4-amino-N--                                                                           1, 170.78  18.02  9.48*                                                                             41.72  4.09*                                       (alpha-                                                                               1/2,                                                                              (153.82-189.96)                                                                        (13.41-21.43)                                                                            (38.83-46.00)                                      methylben-                                                                            1/2,                                                                              [15.32]  [6.02]   [13.67]                                            zyl)-ben-                                                                      amide                                                                          Pheno- 1/2,                                                                              69.01   21.78  3.17                                                                              13.17  5.24                                        barbital                                                                              1, (62.84-72.89)                                                                          (14.99-25.52)                                                                             (5.87-15.93)                                             1  [24.67] [14.98]    [5.93]                                            Phenytoin                                                                             2,2                                                                               65.46    9.50  6.89                                                                              No protection                                                                         <0.22                                                 (52.49-72.11)                                                                           (8.13-10.44)                                                                            up to 300                                                    [15.23] [13.67]                                                      Mephenytoin                                                                           1/4,                                                                              153.82  60.50  2.54                                                                              30.45  5.05                                               1/2,                                                                              (132-178.73)                                                                           (49.45-70.25)                                                                            (19.67-39.47)                                             1/2                                                                                [9.22]  [8.01]    [4.76]                                            Carbamaz-                                                                             1/4,                                                                              71.56    8.81  8.12                                                                              No protection                                                                         <0.72                                       epine  1/4                                                                                (45.91-134.79)                                                                         (5.45-14.09)                                                                            up to 100                                                     [4.77]  [3.62]                                                      __________________________________________________________________________      () 95% Confidence interval                                                     [ ] Slope, regression line                                                     *Protection Index (P.I.) = TD50/ED50                                     

As can be seen from the above table, 4-amino-N-(alpha-methylbenzyl)-benzamide has an ED50 of 18.02 mg/kg as measured against maximal electroshock seizures following intraperitoneal (ip) administration in mice and an ED50 of 41.72 mg/kg against SC metrozole. The compound shows a similar level of activity in these tests in rats. The rotorod toxicity of the compound as measured following ip administration in mice is TD50=170.78 mg/kg. This produces a protective index (PI=TD50/ED50) of 9.48 for ip administration in mice. This PI is higher than that for four standard drugs commonly used in anti-convulsant therapy: phenobarbital, PI=3.17; phenytoin, PI=6.89; mephenytoin, PI=2.54; and carbamazepine, PI=8.12.

                                      TABLE 2.                                     __________________________________________________________________________     Profile of Acute Neurotoxicity of 4-amino-N--(alpha-                           methylbenzyl)-benzamide and Some Prototype Anti-                               epileptic Drugs in Mice                                                                                                               HD50                    ADD No.                                                                               Profile of Acute Toxicity, Mice l.p.            LD50*                   or Name                                                                               1 × TD50 2 × TD50    4 × TD50 (mg/kg)                 __________________________________________________________________________     4-amino-                                                                              (171 mg/kg) At 30                                                                             (342 mg/kg) Both animals had                                                                     (684 mg/kg) At 1                                                                              461.76                  N--(alpha-                                                                            min, both animals                                                                             decreased motor activity, atax-                                                                  both animals had                                                                              (421.03-495.04)         methyl-                                                                               had decreased motor                                                                           ia, rotorod toxicity, muscle re-                                                                 creased motor activity,                                                                       [20.85]                 benzyl)-                                                                              activity and ataxia;                                                                          laxation, cyanosis, and decreased                                                                ataxia, rotorod                                                                               718.18 y,               benzamide                                                                             one was spastic, toxic                                                                        respiration at 30 min; one had                                                                   sedation, ptosis,                                                                             (691.28-742.98)                by rotorod test, cyanotic,                                                                    spasticity, sedation, ptosis,                                                                    relaxation, loss of                                                                           [36.24]                        and had decreased respira-                                                                    muscle relaxation and loss of                                                                    ing reflex, decreased                         tion. Both animals appeared                                                                   righting reflex at 1 hr. This                                                                    respiration and cyanosis;                     normal by 8 hrs.                                                                              animal regained righting re-                                                                     one was spastic. At 6 hrs                                    flex at 4 hrs. Both animals ap-                                                                  one was hypnotic, analgesic                                  peared normal at 24 hrs.                                                                         and anesthetized; at 24 hrs                                                    this animal was dead while                                                     the other animal appeared                                                      normal.                                Pheno- (70 mg/kg) Ataxia with in-                                                                    (140 mg/kg) Ataxia with increased                                                                (260 mg/kg) Ataxis with                                                                       135.45                  barbital                                                                              creased motor activity fol-                                                                   motor activity followed by loss of                                                               creased motor activity                                                                        (114.90-177.42)                lowed by sedation and                                                                         righting reflex, tremors, reflex                                                                 lowed by loss of                                                                               [8.41]g                       ptosis with one of two                                                                        scratching, ptosis, sedation and                                                                 reflex, tremors,                                                                              264.70                         animals toxic by the                                                                          some respiratory depression. Both                                                                scratching, ptosis,                                                                           (241.55-285.52)                rotorod test up to 4 hrs.                                                                     mice regained their righting re-                                                                 anesthesia with                                                                               [15.95]ic                                     flex at 2 hrs and one was still                                                                  and respiratory depression.                                  toxic by the rotorod test at 8 hrs                                                               Both animals became cold to                                                    touch and cyanotic with in-                                                    creased respiratory depres-                                                    sion with death at approxi-                                                    mately 3 and 6 hrs.                    Phenytoin                                                                             (65 mg/kg) Increased motor                                                                    (130 mg/kg) Mild ataxia, periods                                                                 (260 mg/kg) Toxic                                                                             178.34 ta-                     activity, slight ataxia,                                                                      of sedation, spasms of arching,                                                                  tions identical to                                                                            (152.93-195.45)                frequent grooming, nervous                                                                    rolling, straub tail, ptosis, &                                                                  At 1 hr righting reflex                                                                       [14.03]                        behavior, & straub tail.                                                                      respiratory depression. Later,                                                                   sent, longer periods of                                                                       229.61                         Animals were not toxic by                                                                     spasms reduced & sedation in-                                                                    dation, & spasms reduced                                                                      (216.44-259.10)                the rotorod test.                                                                             creased. At 24 hrs, toxicity                                                                     head twisting &                                                                               [15.89]inat-                                  present, other symptoms less pro-                                                                ed movements; respiratory                                    nounced.          depression continued, mice                                                     dead at 24 hrs.                        Mepheny-                                                                              (154 mg/kg) Both animals                                                                      (308 mg/kg) Both animals                                                                         (616 mg/kg) Both                                                                              405.97                  toin   were ataxic and had de-                                                                       were ataxic and toxic by                                                                         had decreased motor                                                                           (366.67-453.67)                creased respiration. Both                                                                     rotorod, had decreased motor                                                                     ity, ataxia, rotorod                                                                          [17.00]                        animals appeared normal                                                                       activity, decreased                                                                              toxicity, sedation,                                                                           567.97                         at 8 hrs.      respiration and cyanosis.                                                                        muscle relaxation                                                                             (504.15-640.18)                               Both animals appeared normal                                                                     cyanosis. These                                                                                [9.80]s,                                     at 24 hrs.        except for anesthesia and                                                      analgesia, were still                                                          present in one animal after                                                    24 hrs. The other animal                                                       was normal at 24 hrs.                  Carba- (70 mg/kg) Mild ataxia                                                                        (140 mg/kg) Sedation                                                                             (280 mg/kg) Marked                                                                            172.24                  mazepine                                                                              with one of two animals                                                                       followed by depressed                                                                            hing legs spread,                                                                             (134.12-197.79)                toxic by the rotorod                                                                          respiration with normal                                                                          respiratory depression,                                                                        [5.92]                        test. Normal within 1 hr.                                                                     respiration and mild                                                                             loss of righting                                                                              628.70                                        ataxia returning at 2 hrs.                                                                       ptosis, hypnosis                                                                              (555.77-707.61)                                                 analgesia, retention of                                                        corneal reflex, righting                                                       reflex regained at 2 hrs.                                                      No deaths at 24                        __________________________________________________________________________                                             hrs.                                    *24-hour period                                                          

                                      TABLE 3.                                     __________________________________________________________________________     Profile of Anti-convulsant Activity of Orally                                  Administered 4-amino-N--(alpha-methylbenzyl)-                                  benzamide and Some Prototype Anti-epileptic                                    Drugs in Mice                                                                         Time                                                                           of Rotorod                                                              ADD No.                                                                               Test                                                                              TD50    ED50 (mg/kg) and PI                                          Substance                                                                             (hrs)                                                                             (mg/kg) MES        sc Met                                            __________________________________________________________________________     4-amino-N--                                                                           2, 271.21  47.66  5.69*                                                                              Maximum 25%                                                                             <0.90*                                   (alpha-                                                                               1/2,                                                                              (239.67-301.27)                                                                        (45.11-50.13)                                                                             Protection at 100                                 methylben-                                                                            1/2                                                                                [9.09  [24.79]    & no protection                                   zyl)-benz-                   at 300                                            amide                                                                          Pheno- 2, 96.78   20.09  4.82                                                                               12.59    7.69                                     barbital                                                                              2,  (79.88-115.00)                                                                        (14.78-31.58)                                                                              (7.99-19.07                                             2   [8.51]  [5.20]     [3.84]                                           Phenytoin                                                                             2, 86.71    9.04  9.59                                                                               No protection up                                                                        <0.29                                           2  (80.39-96.09)                                                                           (7.39-10.62)                                                                             to 300                                                      [13.01]  [6.28]                                                      Mephenytoin                                                                           4, 353.93  65.87  5.37                                                                               36.31    9.75                                            2, (291.31-412.36)                                                                        (63.80-68.25)                                                                             (25.79-48.25)                                            1/2                                                                                [6.58] [28.81]     [3.60]                                           Carbamaz-                                                                             1/2,                                                                              217.21  15.44  14.06                                                                              48.07    4.52                                     pine   1/2,                                                                              (131.49-270.11)                                                                        (12.44-17.31)                                                                             (40.75-57.35)                                            2   [3.47]  [9.07]     [5.50]                                           __________________________________________________________________________      () 95% Confidence Interval                                                     [ ] Slope, regression line                                                     *Protective Index (P.I.) = TD50/ED50                                     

                                      TABLE 4.                                     __________________________________________________________________________     Anti-convulsant Potency of Intraperitoneally                                   Administered 4-amino-N--(alpha-methylbenzyl)-                                  benzamide and Some Prototype Anti-epileptic Drugs                              As Measured by Ability to Prevent Chemically-                                  Induced Seizures in Mice                                                              Time                                                                           of Rotorod                                                              ADD No.                                                                               Test                                                                              TD50  ED50 (mg/kg) and PI                                            Substance                                                                             (hrs)                                                                             (mg/kg)                                                                              sc Met     Bicuculline Picrotoxin  Strychnine                  __________________________________________________________________________     4-amino-N--                                                                           1/2,                                                                              170.78                                                                               41.72  4.09*                                                                              39.07   4.37*                                                                              191.23  0.89*                                                                              Maximum                                                                                <0.85*              (alpha-                                                                               1/2,                                                                              (153.82-                                                                             (38.83-46.00)                                                                             (20.27-61.57)                                                                               (84.48-724.71)                                                                            tection of                  methylben-                                                                            1/2                                                                               189.96)                                                                              [13.67]     [2.38]      [1.73]     37.5% at 170;               zyl)-benz-                                                                               [15.32]                                  25% at 200                  amide                                                                          Pheno- 1/2,                                                                              69.01 13.17  5.24                                                                               37.72   1.83                                                                               27.51   2.51                                                                               95.30   0.72                barbital                                                                              1  (62.84-                                                                               (5.87-15.93)                                                                             (26.49-47.39)                                                                              (20.88-34.82)                                                                              (91.31-99.52)                         72.89)                                                                                [5.93]     [4.07]      [4.79]     [18.51]                               [24.67]                                                              Phenytoin                                                                             2, 65.46 No pro-                                                                               <0.22                                                                              No pro- <0.65                                                                              No pro- <0.65                                                                              Max.                               2  (52.49-                                                                              tection up to                                                                             tection up to                                                                              tection up to                                                                              Prot. 50%                             72.11)                                                                               100        100         100         at 55-100                             [15.23]                                                              Mepheny-                                                                              1/4,                                                                              153.82                                                                               30.45  5.05                                                                               124.14  1.24                                                                               100.96  1.52                                                                               Max. Prot.                  toin   1/2                                                                               (132.86-                                                                             (19.67-39.47)                                                                              (84.10-188.49)                                                                             (79.34-122.92)                                                                            50% at 70-                            178.73)                                                                               [4.76]     [1.99]      [7.43]     150                                   [9.22]                                                               Carbamaz-                                                                             1/4,                                                                              71.56 No protection                                                                         <0.72                                                                              100 mg/kg                                                                              <0.72                                                                              37.20   1.92                                                                               78.83   0.91                epine  1/4                                                                               (45.91-                                                                              up to 100  1/2 protected                                                                              (25.32-59.69)                                                                               (39.39-132.03)                       134.79)                       [3.86]      [2.85]                               [4.77]                                                               __________________________________________________________________________      () Confidence Levels, 95%                                                      [ ] Slope, regression line                                                     *Protective Index (P.I) = TD50/ED50                                      

                                      TABLE 5.                                     __________________________________________________________________________     Profile of Anti-convulsant Activity of Orally                                  Administered 4-amino-N--(alpha-methylbenzyl)-                                  benzamide and Some Prototype Anti-epileptic                                    Drugs in Mice                                                                         Time                                                                           of  Rotorod                                                             ADD No.                                                                               Test                                                                               TD50     ED50 (mg/kg) and PI                                        Substance                                                                             (hrs.)                                                                             (mg/kg)  MES        sc Met                                          __________________________________________________________________________     4-amino-N--                                                                           2,  170.08   16.11  10.56*                                                                             Max. Prot.                                                                             *                                       (alpha-                                                                               1,  (129.74-205.75)                                                                         (15.08-17.46)                                                                             50% at 300;                                     methyl-                                                                               1    [5.66]  [19.24]    1 of 8 animals,                                 benzyl)-                       recurring clonic                                benzamide                      seizures**                                      Pheno- 1/2,                                                                               61.09     9.14  6.68                                                                               11.55   5.29                                    barbital                                                                              5,  (43.72-95.85)                                                                            (7.58-11.86)                                                                              (7.74-15.00)                                          5    [3.00]   [4.12]     [4.08]                                         Phenytoin                                                                             1/2,                                                                               >3000    29.82  >100                                                                               No pro-                                                4            (21.92-38.91)                                                                             tection up to                                                        [2.82]    800                                             Mepheny-                                                                              4,  85.73    18.10  4.74                                                                               21.65   3.96                                    toin   2,  (69.88-93.76)                                                                           (14.02-24.91)                                                                             (17.99-25.77)                                          4   [13.47]   [3.60]     [8.43]                                         Carbamaz-                                                                             2,  813.06    8.50  95.65                                                                              Variable                                        epine  1    (488.76-1233.87)                                                                        (3.39-10.53)                                                          [6.06]   [4.50]                                                    __________________________________________________________________________      () Confidence Levels, 95%                                                      [ ] Slope, regression line                                                     *Protective Index (P.I.) = TD50/ED50                                           **After intraperitoneal administration, 150 mg/kg, 8/8 protected vs sc Me      test and 6/8 toxic; 75 mg/kg, 4/8 protected vs sc Met test and 2/8 toxic.

                                      TABLE 6.                                     __________________________________________________________________________     Profile of Anti-convulsant Activity of Intraperitoneally                       Administered 4-amino-N--(alpha-methylbenzyl)-benzamide and                     Some Prototype Anti-epileptic Drugs in Mice                                           Time                                                                           of Rotorod                                                              ADD No.                                                                               Test                                                                              TD50 ED50 (mg/kg) and PI                                             Substance                                                                             (hrs)                                                                             (mg/kg)                                                                             MES       sc Met     Bicuculline                                                                             Picrotoxin                                                                              Strychnine               __________________________________________________________________________     4-amino-N--                                                                           1, 170.78                                                                              18.02  9.48*                                                                             41.72  4.09*                                                                              39.07                                                                               4.37*                                                                              191.23                                                                              0.89*                                                                              Max Prot.                (alpha-                                                                               1/2,                                                                              (153.82-                                                                            (13.41-21.43)                                                                            (38.46.00) (20.27-  (84.48-  37.5% at                 methylben-                                                                            1/2                                                                               189.96)                                                                              [6.02]   [13.67]    61.57)   724.71)  170                      zyl)-ben- [15.32]                   [2.38]   [1.73]                            Pheno- 1/2,                                                                              69.01                                                                               21.78  3.17                                                                              13.17  5.24                                                                               37.72                                                                               1.83                                                                               27.51                                                                               2.51                                                                               95.30                    barbital                                                                              1, (62.84-                                                                             (14.99-25.52)                                                                             (5.87-15.93)                                                                             (26.49-  (20.88-  (91.31-                         1  72.89)                                                                              [14.98]    [5.93]    47.39)   34.82)   99.52)                             [24.67]                   [4.07]   [4.79]   [18.51]                  Phenytoin                                                                             2, 65.46                                                                                9.50  6.89                                                                              No protect-                                                                           <0.22                                                                              No pro-                                                                             <0.65                                                                              No pro-                                                                             <0.65                                                                              Max. Prot.                      2  (52.49-                                                                              (8.13-10.44)                                                                            ion up to  tection  tection  50% at                             72.11)                                                                              [13.66]   300        up to 100                                                                               up to 100                                                                               55-100                             [15.23]                                                              Mepheny-                                                                              1/4,                                                                              153.82                                                                              60.50  2.54                                                                              30.45  5.05                                                                               12.414                                                                              1.24                                                                               100.96                                                                              1.52                                                                               Max. Prot.               toin   1/2                                                                               (132.86-                                                                            (49.45-70.25)                                                                            (19.67-39.47)                                                                             (84.10-  (79.34-  50% at                             178.73)                                                                              [8.01]    [4.76]    188.49)  122.92)  70-150                             [9.22]                    [1.99]   [7.43]                            Carbamaz-                                                                             1/4,                                                                              71.56                                                                                8.81  8.12                                                                              No protect-                                                                           <0.72                                                                              Max.     37.20                                                                               1.92                                                                               78.83 0.91               epine  1/4                                                                               (45.91-                                                                              (5.45-14.09)                                                                            ion up to  Prot.    (25.32-  (39.39-                            134.79)                                                                              [3.62]   100        62.5% at 59.69)   132.03)                            [4.77]                    50-130   [3.86]   [2.85]                   __________________________________________________________________________      () 95% Confidence Interval                                                     [ ] Slope, regressling line                                                    *Protective Index (P.I.) = TD50/ED50                                     

                                      TABLE 7.                                     __________________________________________________________________________     Profile of Anti-convulsant Activity of Orally Administered                     4-amino-N--(alpha-methylbenzyl)-benzamide and Some Prototype                   Anti-epileptic Drugs in Mice and Rats                                                Time                                                                           of Test         MES-ED50              sc Met-ED50                        ADD No.                                                                              (hrs) Rotorod TD50                                                                             (mg/kg)               (mg/kg)                            Substance                                                                            Mice                                                                              Rats                                                                              Mice Rats Mice       Rats       Mice      Rats                     __________________________________________________________________________     4-amino-                                                                             2, 2, 271.21                                                                              170.08                                                                              47.66  5.69*                                                                              16.11  10.56*                                                                             Maximum   Max. Prot.               N--(alpha-                                                                           1/2,                                                                              1, (239.67-                                                                            (129.74-                                                                            (45.11-50.13)                                                                             (15.08-17.46)                                                                             25% Pro-  50% at 300               methylben-                                                                           1/2                                                                               1  301.27)                                                                             205.75)                                                                             [24.79]    [19.24]    tection at                                                                               1 of 8 ani-              zyl)-benzy- [9.09]                                                                              [5.66]                     100 & No  mals recur-              amide                                       Prot. at 300                                                                             ring clon-                                                                     ic seizures              Pheno-                                                                               2, 1/2,                                                                              96.78                                                                               61.09                                                                               20.09  4.82                                                                                9.14  6.68                                                                                12.59                                                                               7.69                                                                               11.58 5.29               barbital                                                                             2, 5, (79.88-                                                                             (43.72-                                                                             (14.78-31.58)                                                                              (7.58-11.86)                                                                             (7.99-    (7.74-                         2  5  115.00)                                                                             95.85)                                                                               [5.20]     [4.12]    19.07)    15.00)                               [8.51]                                                                              [3.00]                     [3.84]    [4.08]                   Phenytoin                                                                            2, 1/2,                                                                              86.71                                                                               >3000                                                                                9.04  9.59                                                                               29.82  >100                                                                               No pro-                                                                              <0.29                                                                              No pro-                        2  4  (80.39-    (7.39-10.62)                                                                             (21.92-38.91)                                                                             tection   tection                              96.09)     [6.28]     [2.82]    up to 300 up to 800                            [13.01]                                                            Mepheny-                                                                             4, 4, 353.93                                                                              85.73                                                                               65.87  5.37                                                                               18.10  4.74                                                                               36.31 9.73                                                                               21.65 3.96               toin  2, 2, (291.31-                                                                            (69.88-                                                                             (63.80-68.25)                                                                             (14.02-24.91)                                                                             (25.79-   (17.99-                        1/2                                                                               4  412.36)                                                                             93.76)                                                                              [28.81]     [3.60]    48.25)    25.77)                               [6.58]                                                                              [13.47]                    [3.60]                                                                               [8.43]                       Carbamaz-                                                                            1/2,                                                                              2, 217.21                                                                              813.06                                                                              15.44  14.06                                                                               8.50  95.65                                                                              48.07 4.51                                                                               Variable                 epine 1/2                                                                               1  (131.49-                                                                            (488.76-                                                                            (12.44-17.31)                                                                              (3.39-10.53)                                                                             (40.75-                                        270.11)                                                                             1233.87)                                                                             [9.07]     [4.50]    57.35)                                         [3.47]                                                                              [6.06]                     [5.50]                             __________________________________________________________________________      () 95% Confidence Interval                                                     [ ] Slope, regression line                                                     *Protective Index (P.I.) = TD50/ED50                                     

                                      TABLE 8.                                     __________________________________________________________________________     Quantitative Toxicity Profile of Intraperitoneally                             Administered 4-amino-N--(alpha-methylbenzyl)-                                  benzamide and Some Prototype Anti-epileptic Drugs                              in Mice                                                                               Time                                                                           of                                                                      Add No.                                                                               Test                                                                              Dose 50 (mg/kg)                                                      Substance                                                                             (hrs)                                                                             Lethality                                                                              Righting Reflex                                                                           Rotorod                                           __________________________________________________________________________     4-amino-N--                                                                           24,                                                                               718.18  461.76  1.56*                                                                             170.78  4.21*                                     (alpha-                                                                               4, (691.28-742.98)                                                                        (421.03-495.04)                                                                           (153.82-189.96)                                   methylben-                                                                            1  [36.24] [20.85]    [15.32]                                           zyl)-benz-                                                                     amide                                                                          Pheno- 24,                                                                               264.70  135.45  1.95                                                                              69.01   3.84                                      barbital                                                                              1, (241.55-285.52)                                                                        (114.90-177.42)                                                                           (62.84-72.89)                                            1/2                                                                               [15.95]  [8.41]    [24.67]                                           Phenytoin                                                                             24,                                                                               229.61  178.34  1.29                                                                              65.46   3.51                                             12,                                                                               (216.44-259.10)                                                                        (152.93-195.45)                                                                           (52.49-72.11)                                            2  [15.89] [14.03]    [15.23]                                           Mephenytoin                                                                           24,                                                                               567.97  405.97  1.40                                                                              153.82  3.69                                             1, (504.15-640.18)                                                                        (366.67-453.67)                                                                           (132.86-178.73)                                          1/4                                                                                [9.80] [17.00]     [9.22]                                           Carbamaz-                                                                             24,                                                                               628.70  172.24  3.65                                                                              71.56   8.79                                      epine  1/4,                                                                              (555.77-707.67)                                                                        (134.12- 197.79)                                                                           (45.96-134.73)                                          1/4                                                                               [10.11]  [5.92]     [4.77]                                           __________________________________________________________________________      () 95% Confidence interval                                                     [ ] Slope, regression line                                                     *Ratio LD50/HD50 or LD50/TD50                                            

For a compound to be useful in man, it should be adequately absorbed after oral administration and should have an adequate margin of safety. The extent of oral absorption can be determined from the ratio of the oral ED50/i.p. (intraperitoneally) ED50 for a desired pharmacologic activity and should be equal to or less than four for adequate absorption. The available data on the MES test in mice yield an oral ED50/i.p. ED50 of 2.65. This suggests that the amide is adequately absorbed in mice after oral administration.

The amide was examined for mutagenic activity in a series of in vitro microbial assays employing Salmonella indicator organisms. The compound was tested directly and in the presence of liver microsomal enzyme preparations from Aroclor-induced rats. The dose range employed for the evaluation was from 1.0 ug to 10,000.0 ug per plate. The results of the tests conducted on the amide in the absence of a metabolic activation system and in the presence of a rat liver activation system were negative. The amide did not exhibit genetic activity in any of the assays conducted in this evaluation.

As can be seen from the above tests, a critical comparative study of the toxicity and anti-convulsant activity of 4-amino-N-(alpha-methylbenzyl)-benzamide has been done by a battery of tests in mice and rats. The results obtained have been compared with similar data for four prototype clinically useful anti-epileptic drugs: phenobarbital, phenytoin, mephenytoin, and carbamazepine. Based on this critical comparison, it has been concluded that:

1. This amide (4-amino-N-(alpha-methylbenzyl)-benzamide) exhibits anti-convulsant activity in mice and rats as measured by ability to abolish the toxic-extensor phase of maximal electroshock seizures and, after intraperitoneal administration, to elevate minimal Metrazol seizure threshold.

2. This amide is also effective against seizures induced by bicuculline; it is effective against picrotoxin seizures in toxic doses, and only partially effective against seizures induced by strychnine (maximum protection of 37.5% with a minimal toxic dose of 170 mg/kg).

3. Observational assessment of toxic symptoms induced by 1TD50, 2TD50's, 4TD50's for this amide indicate the following are most prominent: decreased motor activity, ataxia, rotorod toxicity, muscle relaxation, and loss of righting reflex. Decreased respiration, cyanosis, and spasticity were also seen in some animals. All surviving animals (one animal given 4TD50's was dead at 24 hours) were free of overt toxic effect within 24 hours after drug administration.

4. Protective indices indicate that this amide has an adequate TD50/ED50 ratio. Intraperitoneal P.I.'s (TD50/ED50) in mice for this amide phenobarbital, phenytoin, mephenytoin, and carbamazepine by the MES test are 9.48, 3.17, 6.89, 2.54, and 812, respectively, and by the sc Met test are 4.09, 5.24, <0.22, 5.05, and <0.72, respectively; after oral administration in rats, the P.I.'s by the MES test are 10.5, 6.68, >100, 4.74, and 95.65, respectively, the only prototype agents with significant P.I.'s by the sc Met test are phenobarbital and mephenytoin (5.29 and 3.96, respectively). Thus, the P.I.'s by the MES test for this amide are within the range of those for the comparative prototype agents.

5. The slopes of the regression lines for this amide by the MES, rotorod, and righting reflex tests resemble those for one or more of the prototype agents. The regression line of this amide by the MES test resembles that for mephenytoin; that by the rotorod test, phenytoin; and that by the righting reflex test, phenytoin and mephenytoin. The slopes of the regression lines by the sc Met and 24-hour lethality tests do not resemble those for any of the prototype agents.

6. The ratio between the oral ED50 and i.p. ED50, determined by the MES test in mice, is 2.65. Thus, it would appear that the test compound is adequately absorbed after oral administration in this species.

7. The results presented show that this amide has significant anti-epileptic potential and the anti-convulsant profile resembles that of phenobarbital and mephenytoin.

Some further compounds of the amino-benzamide type contemplated within this invention are the following:

1. 4-amino-N-phenyl-benzamide; [4-aminobenzanilide]

2. 4-amino-N-cyclohexyl-benzamide

3. 4-amino-N-amyl-benzamide; [4-amino-N-(n-pentyl)-benzamide]

4. 4-amino-N-benzyl-benzamide

5. 4-amino-N,N-(di-n-propyl)-benzamide

6. 4-amino-N-(n-hexyl)-benzamide

7. 4-amino-N-(n-butyl)-benzamide

8. 4-amino-N-(N-methylbenzyl)-benzamide ##STR3##

SYNTHESIS

These compounds were made by the same 2-step sequence described for 4-amino-N-(alpha-methylbenzyl)-benzamide. A list of the appropriate acid chloride and amine for each compound is given below:

    ______________________________________                                         No.   Acid Chloride    Amine                                                   ______________________________________                                         1     4-nitrobenzoylchloride                                                                          aniline                                                 2     "                cyclohexylamine                                         3     "                amylamine                                               4     "                benzylamine                                             5     "                di-n-propylamine                                        6     "                n-hexylamine                                            7     "                n-butylamine                                            8     "                N--methylbenzylamine                                    9     3-nitrobenzoylchloride                                                                          alpha-methylbenzylamine                                 10    2-nitrobenzoylchloride                                                                          alpha-methylbenzylamine                                 ______________________________________                                    

BIOLOGICAL DATA ON SIMILAR COMPOUNDS

The available activity data on these similar amino-benzamides is given below: ##STR4##

                                      TABLE 9.                                     __________________________________________________________________________                              Anti-convulsant Data (mg/kg)*                         No.                                                                               R.sup.1       R.sup.2 TD50                                                                               ED50 MES                                                                             ED50 sc Met                                 __________________________________________________________________________     1  H             --C.sub.6 H.sub.5                                                                      111.30                                                                             50.54 59.11                                       2  H             --C.sub.6 H.sub.12                                                                     188.56                                                                             67.18 123.34                                      3  H             --(CH.sub.2).sub.4 CH.sub.3                                                             68.55                                                                             42.98 57.27                                       4  H             --CH.sub.2 --C.sub.6 H.sub.5                                                            83.44                                                                             23.30 --                                          5  --CH.sub.2 CH.sub.2 CH.sub.3                                                                 --CH.sub.2 CH.sub.2 CH.sub.3                                                           115.54                                                                             61.66 --                                          6  H             --(CH.sub.2).sub.5 --CH.sub.3                                                          Activity at 100 mg/kg or less                         7  H             --(CH.sub.2).sub.3 --CH.sub.3                                                          Activity at 100 mg/kg or less                         8  CH.sub.3      --CH.sub.2 --C.sub.6 --H.sub.5                                                         Activity at 100 mg/kg or less                         9  3-amino-N--(alpha-methyl-                                                      benzyl)benzamide-m-amino-                                                      N--(alpha-methylbenzyl)-                                                       benzamide             Activity at 100 mg/kg or less                         10 2-amino-N--(alpha-methyl-                                                      benzyl)benzamide-o-amino-                                                      N--(alpha-methylbenzyl)-                                                       benzamide             Activity at 100-300 mg/kg                             __________________________________________________________________________      *all data obtained by i.p. administration in mice                        

As can be seen from Table 9, these other amino-benzamide compounds have similar activities to those of 4-amino-N-(alpha-methylbenzyl)-benzamide. It is felt that they may be substituted as indicated above for the first compound and should have similar toxicity and absorption levels. It is, therefore, apparent from the above Tables and Examples that these compounds may be substituted for one another and used in similar manner.

With regard to dosage levels for human beings, it can be seen from the differences in dosage levels per kilogram weight used for mice versus that used for rats that when dealing with human beings it would be necessary to determine this level by testing over a wide range using very small amounts worked up to larger amounts and the level is then determined by the side-effects and its activity.

As this invention may be embodied in several forms without departing from the spirit or essential characteristics thereof, the present embodiment is, therefore, illustrative and not restrictive, since the scope of the invention is defined by the appended claims rather than by the description preceding them, and all changes that fall within the mete and bounds of the claims or that form their functional as well as conjointly cooperative equivalent are, therefore, intended to be embraced by those claims. 

I claim:
 1. A method of treating epilepsy and other convulsive disorders comprising administering an N-substituted amino-benzamide selected from the group consisting of 4-amino-N-phenyl-benzamide, 4-amino-N-cyclohexyl-benzamide, 4-amino-N-amyl-benzamide, 4-amino-N-benzyl-benzamide, 4-amino-N,N-(di-n-propyl)-benzamide, 4-amino-N-(n-hexyl)-benzamide, 4-amino-N-(n-butyl)-benzamide, 3-amino-N-(alpha-methylbenzyl)-benzamide, 2-amino-N-(alpha-methylbenzyl)-benzamide, and 4-amino-N-(alpha-methylbenzyl)-benzamide.
 2. The method of claim 1 wherein the benzamide is administered subcutaneously in an effective amount.
 3. The method of claim 1 wherein the benzamide is administered intraperitoneally in an effective amount.
 4. The method of claim 1 wherein the benzamide is administered orally in an effective amount.
 5. 4-amino-N-(alpha-methylbenzyl)-benzamide. 